A cellulose-producing strain isolated from vinegar culture, preservation number TCCC 10025 in the Tianjin university of science & technology’s center of culture collection, was identified as Gluconacetobacter xylinus based on morphological character, traditional physiological and biochemical method and 16S rDNA complete sequencing analysis. The composition of medium and fermentation conditions was studied. When the initial pH of the medium was 6.0, which contained 70g/L sucrose, 9g/L yeast extract, 9g/L tryptone, trace amount of Ca²⁺, SiO₃^2- and Mg²⁺, and the strain was cultivated at 28^oC, the amount of bacterial cellulose produced by Gluconacetobacter xylinus TCCC 10025 was the highest and reached to 9.214±0.053 g/L. The film produced by Gluconacetobacter xylinus TCCC 10025 was turn out to be bacterial cellulose by the SEM and FT-IR analysis.